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Biomedical subjects

Zhong Zhong

Publications and source records attributed to Zhong Zhong.

13 recordsLinked to original sources

Reliability of diffraction enhanced imaging for assessment of cartilage lesions, ex vivo.

OBJECTIVE: The assessment of articular cartilage integrity is of value for the detection of early degenerative joint disease in both the clinical and the research settings. It was the purpose of this study to determine the accuracy and reliability of identifying articular cartilage defects through Diffraction Enhanced Imaging (DEI), a high contrast radiographic imaging technique. DEI provides two new sources of image contrast to radiography: refraction and scatter rejection, besides the absorption of conventional radiography. DESIGN: Cadaveric tali were DEI imaged in the anterior-posterior position at the National Synchrotron Light Source. Two independent observers provided gross score evaluations (on a five point scale) of the trochlear surfaces. The DEI image of each trochlear surface was then graded (on a five point scale) by two additional independent observers who were blinded with regard to the gross evaluation of the articular surfaces. Inter-observer agreement for DEI grades was assessed with the weighted kappa statistic. Correlation of diffraction enhanced image score to the gross score was assessed with Spearman correlation coefficient. RESULTS: The defects of articular cartilage of talar trochleae could be visualized through DEI. The Spearman correlation of gross grades with DEI grades on the 165 talar regions for observers 1 and 2 were 0.91 and 0.91, respectively. The overall weighted kappa value for inter-observer agreement was 0.93, thus considered high agreement. CONCLUSIONS: DEI is accurate and reliable for detection of articular cartilage defects ex vivo. Even early stages of degeneration of cartilage can be visualized with this high contrast technique. Future studies will focus on the application of DEI to the identification of such lesions in vivo.

Aged↗

Application of division arrest technology to cell-based HTS: comparison with frozen and fresh cells.

Cell-based functional assays are becoming popular in many HTS laboratories because of recent advances in detection and automation technologies. However, the supply of large amounts of live cells with consistent cellular response for day-to-day screening operations over several days/weeks is a tremendous challenge. The high cost of cell culture, labor-intensive nature of the work, and inherent variability in cellular responses from time to time tend to be prohibitive for extensive applications of cell-based assays in HTS. We therefore tested division-arrested cells that were prepared in a single batch and frozen at -80 degrees C before use in several cell-based assays and in a robotic screening campaign. Chinese hamster ovary cells expressing a Gq-coupled receptor were analyzed for the agonist-induced intracellular Ca2+ response measured on a fluorescent imaging plate reader. In this case, the division-arrested cells showed consistent agonist-induced intracellular Ca2+ concentration response as reflected by signal-to-basal ratio and EC50 even 48 h after cell plating. In comparison, the responses from untreated frozen cells and fresh cells declined significantly approximately 30 h after cell plating. In other cell-based assays tested (cyclic AMP assay, reporter gene beta-lactamase assay, and ion-channel assay), the division-arrested cells performed as well as frozen, or fresh cells. We thus conclude that the use of alternate strategies such as frozen cells or division-arrested cells may alleviate the need for several batches of cell plating each day during HTS while maintaining the desired robotic throughput and assay quality.

Animals↗

Improving consistency of cell-based assays by using division-arrested cells.

In this article we describe the use of division-arrested cells for cell-based assays designed for high-throughput screening. Cells are the most critical and variable reagent for cell-based high-throughput screening. The robustness of robotic screening depends on the quality and consistency of cell reagents. We demonstrate that for most cell types commonly used for high-throughput screening, cells can be irreversibly division-arrested by mitomycin C treatment at doses that cause no apparent toxicity or obvious change to the cell signaling properties we measured. Our data also suggest that division-arrested cells perform favorably compared to regular growing cells in reporter and calcium flux assays, two platforms most commonly used in robotic screening. Division arrest technology effectively uncouples the process of cell production from robotic screening and brings the convenience of having quality-approved cell reagent on demand for cell-based high-throughput screening.

Animals↗

Improved image contrast of calcifications in breast tissue specimens using diffraction enhanced imaging.

The contrast of calcifications in images of breast tissue specimens using a synchrotron-based diffraction enhanced imaging (DEI) apparatus has been measured and is compared to the contrast in images acquired using a conventional synchrotron-based radiographic imaging modality. DEI is an imaging modality which derives image contrast from x-ray absorption, refraction and small-angle scatter-rejection (extinction), unlike conventional radiographic techniques, which can only derive contrast from absorption. DEI is accomplished by inserting an analyser crystal in the beam path between the sample and the detector. Two of the three breast tissue specimens contained calcifications associated with cancer, while a third contained benign calcifications. Results of the image analysis indicate that the DEI contrast of images taken with the analyser crystal tuned to the peak of its rocking curve, was as much as 19 times that of the conventional radiograph, with an average of 5.5 for all calcifications. This improved image contrast for even near-pixel-size calcifications suggests potential utility for DEI in breast imaging.

Breast Diseases↗

Multiple-image radiography.

Conventional radiography produces a single image of an object by measuring the attenuation of an x-ray beam passing through it. When imaging weakly absorbing tissues, x-ray attenuation may be a suboptimal signature of disease-related information. In this paper we describe a new phase-sensitive imaging method, called multiple-image radiography (MIR), which is an improvement on a prior technique called diffraction-enhanced imaging (DEI). This paper elaborates on our initial presentation of the idea in Wernick et al (2002 Proc. Int. Symp. Biomed. Imaging pp 129-32). MIR simultaneously produces several images from a set of measurements made with a single x-ray beam. Specifically, MIR yields three images depicting separately the effects of refraction, ultra-small-angle scatter and attenuation by the object. All three images have good contrast, in part because they are virtually immune from degradation due to scatter at higher angles. MIR also yields a very comprehensive object description, consisting of the angular intensity spectrum of a transmitted x-ray beam at every image pixel, within a narrow angular range. Our experiments are based on data acquired using a synchrotron light source; however, in preparation for more practical implementations using conventional x-ray sources, we develop and evaluate algorithms designed for Poisson noise, which is characteristic of photon-limited imaging. The results suggest that MIR is capable of operating at low photon count levels, therefore the method shows promise for use with conventional x-ray sources. The results also show that, in addition to producing new types of object descriptions, MIR produces substantially more accurate images than its predecessor, DEI. MIR results are shown in the form of planar images of a phantom and a biological specimen. A preliminary demonstration of the use of MIR for computed tomography is also presented.

Absorptiometry, Photon↗

Measurement of image contrast using diffraction enhanced imaging.

Refraction contrast of simple objects obtained using diffraction enhanced imaging (DEI) was studied and compared to conventional radiographic contrast. Lucite cylinders and nylon wires were imaged using monochromatic synchrotron radiation at the National Synchrotron Light Source (http://nslsweb. nsls.bnl.gov/nsls/Default.htm) at the Brookhaven National Laboratory. The DEI images were obtained by placing a silicon analyser crystal tuned to the [333] diffraction plane in the beam path between the sample and the detector. To compare the DEI images with conventional radiographic images requires a consistent definition of refraction and absorption contrast. Conventional definitions of contrast favour conventional radiography and DEI contrast is defined to emphasize the specific characteristics of DEI. The proposed definitions were then used to find the DEI gain (the ratio of the DEI contrast with respect to the conventional image contrast). The results presented here show that the DEI gain is consistently greater than 1, indicating that DEI provides more contrast information than conventional radiography.

Computer Simulation↗

Radiography of rabbit articular cartilage with diffraction-enhanced imaging.

Articular cartilage of synovial joints is not visible with conventional X-ray imaging. Hence, the gradual degeneration and destruction of articular cartilage, which is characteristic of degenerative joint diseases, is only detected at a late stage when the cartilage is lost and the joint space that it once occupied narrows. The development of an X-ray imaging technique that could detect both the degenerative cartilage and bone features of joint diseases is of special interest. Here we show, for the first time, that a high-contrast imaging technique, diffraction-enhanced X-ray imaging (DEI), allows the visualization of articular cartilage of both disarticulated and articulated rabbit knee joints. Furthermore, a single cartilage lesion can be visualized within an intact joint. The results suggest that DEI has the potential to be of use in the study of cartilage degeneration.

Animals↗

Radiography of soft tissue of the foot and ankle with diffraction enhanced imaging.

Non-calcified tissues, including tendons, ligaments, adipose tissue and cartilage, are not visible, for any practical purposes, with conventional X-ray imaging. Therefore, any pathological changes in these tissues generally necessitate detection through magnetic resonance imaging or ultrasound technology. Until recently the development of an X-ray imaging technique that could detect both bone and soft tissues seemed unrealistic. However, the introduction of diffraction enhanced X-ray imaging (DEI) which is capable of rendering images with absorption, refraction and scatter rejection qualities has allowed detection of specific soft tissues based on small differences in tissue densities. Here we show for the first time that DEI allows high contrast imaging of soft tissues, including ligaments, tendons and adipose tissue, of the human foot and ankle.

Adipose Tissue↗

New pressure flow cell to monitor BaSO4 precipitation using synchrotron in situ angle-dispersive X-ray diffraction.

A flow cell has been commissioned to monitor in situ precipitation reactions under non-ambient conditions. The majority of high-pressure systems use anvils and presses to obtain high pressures around a small reaction area; however, this prototype is unique in that solutions may be examined as they flow through the cell under pressure. The cell is made of single-crystal silicon, which is capable of withstanding the high pressures created by liquid flow within the cell. With the capability of reaching pressures of up to 4 x 10(7) Pa, the cell is ideal for work on geological and oilfield systems. Here it is used to examine the formation of barium sulfate scale in situ under non-ambient conditions using angle-dispersive XRD on beamline X17b1 at the NSLS.

Journal Article↗

Flavonol glycosides from Lysimachia capillipes.

Two new compounds, capilliposide I and capilliposide II, together with a known compound quercetin-3-O-(2,6-di-alpha-L-rhamnopyranosyl)-beta-D-galactopyranoside were isolated from the extracts of the whole herbs of Lysimachia capillipes. Their structures were established through their spectral data and chemical properties.

Chemistry Techniques, Analytical↗

Simultaneous intracellular calcium and sodium flux imaging in human vanilloid receptor 1 (VR1)-transfected human embryonic kidney cells: a method to resolve ionic dependence of VR1-mediated cell death.

The vanilloid receptor 1 (VR1) is a ligand-gated, nonselective cation channel important for the sensory processing of painful stimuli. Activation of VR1 leads to increases in intracellular concentrations of calcium and sodium. Prolonged activation of VR1 in mammalian expression systems leads to cell death. The mechanism of VR1-mediated toxicity may have relevance to pathophysiological processes that can occur in neurons. Therefore, we have evaluated the relative contributions of intracellular calcium and sodium changes to VR1-mediated toxicity in human embryonic kidney 293 cells stably transfected with the human VR1 channel. The data demonstrate that VR1 receptor agonists capsaicin and resiniferatoxin lead to a sustained increase in intracellular calcium and sodium in a concentration-dependent manner, followed by cell death. Pretreatment with VR1 receptor antagonists capsazepine or ruthenium red block both the calcium and sodium responses to agonists, and block agonist-induced cell death in a concentration-dependent manner. However, addition of antagonists several minutes after agonists selectively reverses the agonist-induced increase in intracellular calcium, but does not reverse the elevated intracellular sodium concentration. Nonetheless, antagonists retain protective efficacy against capsaicin toxicity when added several minutes after capsaicin, conditions in which the cells still manifest elevated intracellular sodium, but not elevated intracellular calcium. In addition, a transient VR1-mediated increase in intracellular calcium that returns to baseline within minutes, induced by a rapid drop in pH, from pH 7.5 to pH 6.3, also does not lead to cell death. Collectively, these data demonstrate that the most important intracellular ionic change for mediating VR1-dependent toxicity is a sustained increase of calcium.

Benzofurans↗

Radiography of soft tissue of the foot and ankle with diffraction enhanced imaging.

Non-calcified tissues, including tendons, ligaments, adipose tissue and cartilage, are not visible, for any practical purposes, with conventional X-ray imaging. Therefore, any pathological changes in these tissues generally necessitate detection through magnetic resonance imaging or ultrasound technology. Until recently the development of an X-ray imaging technique that could detect both bone and soft tissues seemed unrealistic. However, the introduction of diffraction enhanced X-ray imaging (DEI) which is capable of rendering images with absorption, refraction and scatter rejection qualities has allowed detection of specific soft tissues based on small differences in tissue densities. Here we show for the first time that DEI allows high contrast imaging of soft tissues, including ligaments, tendons and adipose tissue, of the human foot and ankle.

Ankle↗

Refraction effects of diffraction-enhanced radiographic imaging: a new look at bone.

The objective of this study was to demonstrate the ability of a novel radiographic technology-diffraction-enhanced imaging-to detect contrast in bone tissue through absorption, refraction, and scatter rejection. Diffraction-enhanced imaging uses a synchrotron x-ray beam to produce images of high contrast by measuring the object's refraction and ultra-small angle scattering of x-rays in addition to the attenuation measured by conventional radiography. We present evidence that diffraction-enhanced imaging provides contrast enhancement at the edges of cortical and cancellous bone and a three-dimensional appearance of trabeculae.

Animals↗